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大黄鱼EBI3基因的克隆与表达分析
霍洁颖,刘小玲,林 蠡,陈新华
0
(华中农业大学水产学院,湖北 武汉 430070;国家海洋局第三海洋研究所海洋生物遗传资源重点实验室,福建 厦门 361000)
摘要:
通过克隆大黄鱼(Larimichthys crocea)的EBI3(Epstein-Barr virus-induced gene 3)基因并对其进行了序列分析.大黄鱼EBI3(LycEBI3)基因开放阅读框长747bp,编码248个氨基酸,存在1个信号肽序列和2个FN3结构域(37~130,145~230).多序列比对发现LycEBI3分子与其他已知EBI3氨基酸水平一致性为27.68%~57.53%.Real-time PCR结果表明,LycEBI3基因在脾脏和血液中表达量最高,且溶藻弧菌(Vibrio alginolyticus)刺激后,大黄鱼头肾和脾脏中LycEBI3基因的转录水平会显著升高,说明LycEBI3基因可能参与了抑制由细菌引起的炎症反应.
关键词:  海洋生物学  大黄鱼  EBI3  分子克隆  实时定量  PCR  表达分析
DOI:10.3969/J.ISSN.2095-4972.2017.03.014
基金项目:国家自然科学基金资助项目(31372556)
Cloning and expression analysis of EBI3 gene in large yellow croaker (Larimichthys crocea)
HUO Jie-ying,LIU Xiao-ling,LIN Li,CHEN Xin-hua
(Department of Aquatic Animal Medicine, College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China;Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, SOA, Xiamen 361005, China)
Abstract:
In this research, we cloned and analyzed the sequence of Epstein-Barr virus-induced gene 3 (EBI3) in large yellow croaker (Larimichthys crocea). The open reading frame of LycEBI3 consists of 747 bp encoding a polypeptide of 248 amino acids. The predicted amino acid sequence showed that LycEBI3 contains a signal peptide of 23 amino acids and 2 Fibronectin type 3 (FN3) (residues 37~130, 145~230) domain that is preserved in lower and higher vertebrates. Multiple sequence alignment showed that the LycEBI3 shares 27.68%~57.53% amino acid sequence identity with other known EBI3. LycEBI3 were constitutively expressed in different organs with the higher expression in spleen and blood. The up regulation of LycEBI3 in spleen and head kidney after bacterial infection indicated its involvement in innate immune regulation of suppressing the inflammation during bacterial infections.
Key words:  marine biology  large yellow croaker  Epstein-Barr virus-induced gene 3 (EBI3)  molecule cloning  Real-time PCR  expression analysis

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